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An Optimised Faecal microRNA Sequencing Pipeline Reveals Fibrosis in Trichuris muris Infection

ABSTRACT: The intestine is a site of diverse functions including digestion, nutrient absorption, immune surveillance, and microbial symbiosis. As such, intestinal homeostasis is vital for overall wellbeing. Faecal microRNAs (miRNAs) offer valuable non-invasive insights into the transcriptional state of the intestine. However, typical faecal miRNA yields and profiles remain incompletely characterised. Here, we develop an optimised protocol for faecal miRNA detection, and describe a reproducible murine faecal miRNA profile across several studies by performing a meta-analysis. By examining faecal miRNA changes during chronic infection with the gastrointestinal helminth, Trichuris muris, we identify the altered expression of miRNAs associated with fibrosis, barrier integrity and wound healing. Fibrosis was confirmed in vivo, suggesting a role for these miRNAs in regulating wound healing during chronic infection where the production of classical wound healing Th2 cytokines are low. Further implementations of this technique can identify novel hypotheses and therapeutic strategies in diverse disease contexts.

ORGANISM(S): Mus musculus

PROVIDER: GSE267555 | GEO | 2024/12/20

REPOSITORIES: GEO

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Project description:The current treatment for Celiac Disease (CD) is adhering to a gluten-free diet (GFD), although its long-term molecular effects are still undescribed. New molecular features detectable in faecal samples may improve and facilitate non-invasive clinical management of CD on GFD. For this purpose, faecal small non-coding RNAs (sncRNAs) and gut microbiome profiles were concomitantly explored in CD subjects in relation to strict (or not) GFD adherence over time. In the present observational study, we performed small RNA and shotgun metagenomic sequencing in stool from 63 treated CD (tCD) subjects and 66 sex- and age-matched healthy controls. tCD included 51 individuals on strict GFD and with negative transglutaminase (TG) serology (tCD-TG-) and 12 symptomatic with not strict/short-time of GFD adherence and positive TG serology (tCD-TG+). Samples from additional 40 adult healthy individuals and from a cohort of 19 untreated paediatric CD subjects and 19 sex/age matched controls were analyzed to further test the outcomes. Several miRNA, other sncRNA (piRNA and tRNA) and microbiota profiles were altered in tCD subjects(adj.p<0.05). Findings were validated in one external group of controls. In tCD-TG-, GFD duration correlated with five miRNA levels (p<0.05): for miR-4533-3p and miR-2681-3p, the longer the diet adherence, the less the expression differed from controls. tCD-TG+ and untreated paediatric CD patients showed a similar miRNA dysregulation. Immune-response, trans-membrane transport and cell death pathways were enriched in targets of identified miRNAs. Bifidobacterium longum, Ruminococcus bicirculans and Haemophilus parainfluenzae abundances shifted (adj. p<0.05) with a progressive reduction of denitrification pathways with GFD length. Integrative analysis highlighted 121 miRNA-bacterial relationships (adj.p<0.05). Specific faecal sncRNA and microbial patterns characterise CD subjects on GFD, reflecting either the long-term effects or the gut inflammatory status, in case of a not strict/short-time adherence. Our findings suggest novel host-microbial interplays and could help the discovery of biomarkers for the clinical monitoring of GFD over time.

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An Optimised Faecal microRNA Sequencing Pipeline Reveals Fibrosis in Trichuris muris Infection

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